Objective To construct a chimeric infectious clone of the fatal virulent strain SDLY 107, containing the gene fragments encoding 2A and 3B proteins of the mild virulent strain SDLY 1, and to establish a reverse genetic system platform for further investigation on virulence of enterovirus 71 strains. Methods The overlap PCR analysis was performed to obtain the gene fragments encoding 2A and 3B pro-teins of the mild virulent strain SDLY 1.The obtained gene fragments were digested and then cloned into a plasmid pMD19-T containing the full-length gene of SDLY 107 strain by using gene replacement strategy. The recombinant RNA was transfected into Vero cells for the preparation of recombinant virus particles.Sev-eral assays including the PCR, indirect immunofluorescence ( IFA) , Western blot and sequencing were per-formed for virus identification.Virus titers were measured by 50%cell culture infective dose ( CCID50 ) and plaque assay.Results The infectious clones of SDLY 107-2A-1 and SDLY 107-3B-1 chimeric virus strains were constructed successfully.Typical cytopathic effect was observed in Vero cells after viral transfection. Identification of the rescued viruses by PCR, IFA, Western blot and sequencing further confirmed the suc-cessful construction of infectious virus strains.The virus titers of SDLY 107-2A-1 and SDLY 107-3B-1 strains detected by CCID50 and plaque assay were 1.25 ×105 PFU/ml and 0.7 ×105 PFU/ml, respectively. Conclusion The chimeric viruses SDLY 107-2A-1 and SDLY 107-3B-1 were rescued successfully, causing cytopathic effects similar to those by using the parental virus strain SDLY 107.This study might pave the way for further investigation on in vitro and in vivo virulence of enterovirus 71 strains.

OBJECTIVETo study the proliferation and differentiation of neural stem cells in the subventricular zone (SVZ) in neonatal rats after bilateral common arteries occlusion.

METHODSNinety-six 3-day-old Sparuge-Dawley rats were randomly divided into two groups: ischemia and control. Rats in the ischemia group were subjected to bilateral common arteries occlusion and the rats in the control group were sham-operated. All rats were administrated with 5-bromodeoxyuridine (BrdU) (50 mg/kg) via intraperitoneal injection. Rats were sacrificed and their brains were removed 1, 4, 7, 10, 14 and 35 days after ischemia. Using brain paraffin sections and immunofluorescence assays, the number of newborn cells in the SVZ was counted. Newborn neural stem cells and oligodendrocytes in the SVZ were observed, and then double marked with BrdU and nestin or osmium tetroxide (O4).

RESULTSThe number of BrdU+ cells (neural stem cells) in the SVZ in the ischemia group was greater than in the control group 4, 7, 10 and 14 days after ischemia, and reached a peak at 4 days after ischemia (253.1+/- 49.3 vs 133.5+/- 17.7; P< 0.01). By 35 days after ischemia, the number of BrdU+/O4+ cells (oligodendrocytes) in the corpus callosum (56.0+/- 7.2 vs 17.0+/- 6.4; P< 0.01), the septal nuclei (45.0+/- 11.9 vs 20.5+/- 5.0; P< 0.01), the striatum (34.5+/- 4.2 vs 14.5+/- 5.8; P< 0.01) and the olfactory bulb (46.5+/- 6.6 vs 23.5+/- 8.4; P< 0.01) in the ischemia group increased significantly as compared to the control group (P< 0.01).

CONCLUSIONSBrain ischemia can activate the proliferation of neural stem cells in the SVZ and promote neural stem cells differentiation into oligodendrocytes. The immature brain may have the capacity for self-repair after ischemic brain injury.

Animals ; Animals, Newborn ; Brain Ischemia ; physiopathology ; therapy ; Bromodeoxyuridine ; metabolism ; Cell Differentiation ; Cell Proliferation ; Cerebral Ventricles ; physiopathology ; Female ; Male ; Neurogenesis ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation

Objective To investigate the effects of in situ venous arterialization on extensive artery obliterans occlusion of the lower extremity. Methods Lumbar sympathetic ganglionectomy and one stage in situ arterialization of the great saphemous vein were performed in 104 ischemic limbs of 88 patients with extensive arterial occlusion. Results Eighty-two of 104 limbs were followed-up from 6 months to over 6 years. The intermittent claudication, night pain improved in all cases, with satisfactory wound healing and no swelling of the lower limbs. Conclusions Arterial blood flow through venous conduit improves and reconstructs the blood circulation of the ischemic limbs.

Objective To Design a medical information system to meet the requirements for informatized medical support by the field medical support institution. Methods J2EE architecture and Java technology were applied to the design of the system, and star topology was used for network deployment. Results The system shared casualty treatment data with servicemen electronic tag and medical commanding information with logistics integrated command and control platform,so that the requirements for medical information were met in the institution.Conclusion The system improves the field medical support institution in medical support ability,treatment rate and efficacy.

BACKGROUNDLaparoendoscopic single-site surgery (LESS) approaches have been reported for treating various kidney and pelvic procedures, and are feasible and effective in selected patients. In this study, we aimed to present the initial experience and evaluate the efficacy of laparoscopic radical prostatectomy performed through a single incision using a multichannel port.

METHODSBetween July 2010 and April 2011, six patients diagnosed with early stage prostate cancer underwent LESS radical prostatectomy (RP) in our institute. A multichannel port was inserted transperitoneally through a 2-cm umbilical incision. Specially articulating and flexible laparoscopic were used. Some technical tricks and points were applied during the operation to overcome the drawbacks and reduce the difficulties of this approach. Two continuous urethrovesical sutures in both sides were performed to complete both lateral aspects of anastomosis. The two ends of the suture threads were fixed by double Lapro-Clips, instead of the difficult knot-tying.

RESULTSTotal operative time was (265 ± 43) minutes. Mean blood loss was (230 ± 65) ml. All cases were completed successfully, without conversion to open surgery or adding additional abdomen ports. No patient required a blood transfusion and no intraoperative complications occurred. The Foley catheter was removed at the 14th day (range 12th - 16th) after surgery. At the 12th week of follow-up, all patients had an undetectable prostate-specific antigen level. Two patients used 2 or 1 pad for continence daily; other patients had achieved good continence.

CONCLUSIONIn selected cases, LESS-RP is feasible and effective; these technic points and the flexible-articulating instruments are helpful to reduce the operation difficulties.

Aged ; Humans ; Laparoscopy ; methods ; Male ; Prostatectomy ; methods

OBJECTIVETo analyze the tendency in the incidence of malignant tumors in nutrition-intervened population of Lin County.

METHODSSubjects who had been enrolled in the General Population Trial, Lin County Nutrition Intervention Trial were followed up and the data of malignant tumors were collected with collaboration of National Cancer Institute/National Institute of Health. Incidences of malignant tumors during and after nutrition intervention were calculated and compared.

RESULTSEsophageal cancer, cardiac cancer, and stomach cancer were constantly the three leading cancers, accounting for 84.06% and 83.74% of the malignant tumors in men and women, respectively. The incidences of lung cancer and liver cancer in men showed increasing tendencies, while similar tendencies were found in those of stomach cancer, colorectal cancer, and liver cancer in women. With the prolonging of the follow-up, the incidences of upper digestive tract cancers declined (except stomach cancer in women).

CONCLUSIONSThe upper digestive tract cancers remain the leading malignant tumors, although their incidences may decline with longer-term follow-up. Prevention and control of tumors in Lin County should still be focused on the esophageal cancer, cardiac cancer, and stomach cancer, as well as lung cancer and liver cancer.

Adult ; Aged ; China ; epidemiology ; Diet ; Feeding Behavior ; Female ; Follow-Up Studies ; Humans ; Incidence ; Male ; Middle Aged ; Neoplasms ; epidemiology ; Prospective Studies ; Risk Factors

OBJECTIVETo explore effect of interleukin 10 (IL-10) gene-modified bone marrow-derived liver stem cells (BDLSCs) transplantation on hepatic inflammatory response and liver regeneration in rats with liver fibrosis.

METHODS50 female Wistar rats were randomly divided into 4 groups: (1) control group: 10 rats were subcutaneously injected with olive oil for 8 weeks; (2) fibrosis groups: 16 rats were subcutaneously injected with carbon tetrachloride (CCl4) for 8 weeks to induce liver fibrosis; (3) BDLSC group: 12 rats were subcutaneously injected with CCl4 for 8 weeks, and were transplanted with 2 x 10(5) BDLSC at week 4; (4) BDLSC/IL-10 group: 12 rats were subcutaneously injected with CCl4 for 8 weeks, and were transplanted with 2 x 10(5) IL-10 gene-modified BDLSC at week 4. IL-10 and tumor necrosis factor alpha (TNFa) in liver tissues were detected by ELISA. HE stained liver tissues were observed under light microscope. The expression of hepatocyte growth factor (HGF) was quantified by real-time RT-PCR, and the expression of proliferating cell nuclear antigen (PCNA) was determined by immunohistochemistry.

RESULTSThe ratio of IL-10/TNFa in fibrosis group (0.05+/-0.01) was lower than that in control group (0.26+/-0.04) (P < 0.01). Transplantation of untreated BDLSCs did not improve the ratio (P > 0.05), however, transplantation of IL-10 modified BDLSCs improved the ratio significantly (P < 0.01). Severe inflammatory response and fibrosis were observed in fibrosis group. Inflammatory response was alleviated to some extent in the BDLSC group, and the histopathology of BDLSC/IL-10 group was not significantly different from that of the control group. Compared to the control group, the expression of HGF mRNA and PCNA protein was increased in the fibrosis group (P < 0.01). The expression of HGF and PCNA was further increased by BDLSCs or IL-10 modified BDLSCs transplantation. Compared to BDLSCs, IL-10 gene-modified BDLSCs were more potent to induce the expression of HGF and PCNA.

CONCLUSIONTransplantation of IL-10 gene-modified BDLSCs can alleviate hepatic inflammatory response and promote liver regeneration in hepatic fibrosis rats.

Adenoviridae ; genetics ; Animals ; Bone Marrow Cells ; cytology ; Cell Proliferation ; Disease Models, Animal ; Female ; Genetic Therapy ; methods ; Hepatocyte Growth Factor ; genetics ; metabolism ; Immunohistochemistry ; Interleukin-10 ; genetics ; Liver ; metabolism ; pathology ; Liver Cirrhosis ; genetics ; pathology ; therapy ; Liver Regeneration ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cell Transplantation ; methods ; Transduction, Genetic ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo evaluate the diagnostic potential of previously published enterovirus (EV) reverse transcription polymerase chain reaction (RT-PCR) assay in detection of EV in CSF samples from children with a diagnosis of aseptic meningitis and to investigate the clinical characteristics of the patients seen in Shandong.

METHODSEV RNA was detected in 187 CSF samples and serum and/or urine samples of a part of patients by RT-PCR and viral culture technique.

RESULTSRT-PCR was positive in all 62 CSF specimens which were positive by cell culture (100%). In addition, 93 of 125 (74.4%) CSF samples negative by cell culture were RT-PCR positive. In 4 of these 93 (4.3%) patients, viral culture of specimens from other sites (serum or urine) was also positive. The sensitivity of CSF RT-PCR based on clinical diagnosis in patients with meningitis of negative bacterial culture results was 82.9% (155/187), which was considerably higher than the sensitivity of CSF virus culture 33.2% (62/187). The results of RT-PCR can be reported within 4 hours, whereas the viral culture of CSF requires 4.6 days for a cytopathic effect to develop. EV meningitis occurred in a sporadic form and in some areas there were outbreaks. The clinical characteristics of 155 patients with EV meningitis were different in different age groups.

CONCLUSIONEV was one of the most common causes of aseptic meningitis in Shandong area. The RT-PCR assay was rapid, sensitive and specific for the diagnosis of EV meningitis and may be a potential tests to shorten hospital stay and reduce the use of antibiotics.

Central Nervous System Infections ; blood ; diagnosis ; urine ; Child ; Child, Preschool ; China ; Enterovirus ; genetics ; isolation & purification ; Enterovirus Infections ; cerebrospinal fluid ; diagnosis ; Female ; HeLa Cells ; Humans ; Infant ; Infant, Newborn ; Male ; RNA, Viral ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

McCune-Albright syndrome is a rare G proteins alpha disorder. The disorder is characterized by polyostotic fibrous dysplasia, sexual precocity and hyperpigmented macules. It is caused due to mutations in the gene Gsalpha that incodes the alpha subunit of the trimeric guanosine triphate-binding protein. There is no specific treatment for this syndrome. Treatment is generally symptomatic. This paper reported three cases of McCune-Albright syndrome and reviewed the relevant literatures regarding to the pathogenesis, pathological features, diagnosis and treatment. All three cases presented with a characteristic triad: polyostotic fibrous dysplasia, sexual precocity and hyperpigmented macules and were thus definitely diagnosed with McCune-Albright syndrome.
Child ; Child, Preschool ; Diagnosis, Differential ; Female ; Fibrous Dysplasia, Polyostotic ; diagnosis ; etiology ; pathology ; therapy ; Humans ; Prognosis

To investigate the changes in bcl-2, bax expression and neuron apoptosis in the hippocampus after the blockade of cervical lymphatics, the model of lymphostatic encephalopathy was established by occluding and removing both the superficial and deep cervical lymph nodes in rats. The animals were sacrificed at 1, 2, 3, 5, 7 and 14 d after operation. H and E staining was used to observe the structure of brain tissues and TUNEL staining was used to detect in situ cell apoptosis in the hippocampus. The expression of bcl-2 and bax in the hippocampus were examined by RT-PCR. The results showed that cerebroedema appeared at day 2 and was most serious at day 5 after the blockade of cervical lymphatics. The number of TUNEL positive cells began to increase at day 2 and reached the maximum at day 5. The expression of bax began to increase at day 1 and reached the maximum at day 2. The expression of bcl-2 began to decrease at day 1 and dropped to the minimum at day 5. The items mentioned above recovered to control level at day 14. These results suggest that lymphostatic encephalopathy following the blockade of cervical lymphatics result in changes in bcl-2 and bax expression in the hippocampus and that apoptosis is the main form of neuron death.
Animals ; Apoptosis ; physiology ; Female ; Hippocampus ; metabolism ; pathology ; Lymph Node Excision ; Lymphatic System ; physiology ; Male ; Neck ; Neurons ; cytology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; bcl-2-Associated X Protein ; genetics ; metabolism